CN103479573B - Preparation methods for polyethylene glycol monomethyl ether-polyester diblock copolymer micelle and drug-loaded micelle - Google Patents

Abstract

The invention provides a preparation method of drug-loaded nano-micelles: adding dropwise a mixture of a small molecule drug and an organic solvent into a polyethylene glycol monomethyl ether-polyester diblock copolymer to obtain a mixed solution; The mixed solution was stirred for the first time while adding ultrapure water dropwise, and after the second stirring was continued, the organic solvent was removed by dialysis and freeze-dried to obtain drug-loaded nano micelles. The present invention also provides a preparation method of polyethylene glycol monomethyl ether-polyester diblock copolymer micelles: dissolving polyethylene glycol monomethyl ether-polyester diblock copolymer in an organic solvent, While stirring for the first time, ultrapure water was added dropwise, and after stirring for the second time, the organic solvent was removed by dialysis and freeze-dried to obtain diblock copolymer micelles. The micelles prepared by this method are easy to operate, under mild conditions, and present a good monodisperse state, which can encapsulate small molecule drugs with poor water solubility, and improve the encapsulation efficiency and solubility of drugs, and the obtained drug-loaded nano-micelles have a stable structure , easy to store.

Description

Preparation of polyethylene glycol monomethyl ether-polyester diblock copolymer micelles and drug-loaded micelles method

technical field

The invention relates to the field of macromolecules, in particular to a preparation method of polyethylene glycol monomethyl ether-polyester diblock copolymer micelles and drug-loaded micelles.

Background technique

Polymer carrier drug is an emerging drug delivery technology with the development of pharmaceutical research, biomaterial science and clinical medicine. Low-molecular-weight drugs have the advantages of high curative effect and convenient use, but they also have great side effects. Generally, low-molecular-weight drugs enter the human body through oral administration or injection, with fast metabolism, short half-life, and lack of selectivity. A polymer carrier drug refers to a polymer that has no pharmacological effect and does not react with the drug itself as a carrier of the drug. A class of drugs obtained on the chain. Among them, the high-molecular compound acts as a delivery system for low-molecular drugs.

Using polymer materials as the carrier of small molecule drugs can increase the action time of drugs, improve the selectivity of drugs, reduce the toxicity of small molecule drugs, and position accurately. Micro- and nano-scale polymer carriers, such as nanomicelles, vesicles and nanoparticles, have been rapidly developed in the near future. Such polymer carriers can effectively disperse drug molecules into them, and utilize various response modes of the carrier. , to achieve drug delivery and controlled release.

Among them, the preparation methods of nanomicelle mainly include: self-assembly method, dialysis method, chemical combination method and electrostatic interaction method, etc., but these methods have disadvantages. The nanomicelles prepared by the self-assembly method only rely on non-covalent bonds such as Van der Waals force, hydrogen bond or electrostatic force to connect the material layers, so the mechanical stability of the nanomicelles is poor and the efficiency is low. The chemical combination method requires suitable functional groups to react, and the selection of polymer materials and small molecule drugs is relatively strict. Dialysis and electrostatic interaction methods are not suitable for large-scale production.

Contents of the invention

The technical problem solved by the present invention is to provide a preparation method of polyethylene glycol monomethyl ether-polyester diblock copolymer micelles and drug-loaded micelles, which is simple to operate and easy to industrialize, and the obtained drug-loaded nano micelles High encapsulation efficiency and stable structure.

The invention provides a preparation method of drug-loaded nano micelles, comprising the following steps:

(A) Add the mixture of small molecule drug and organic solvent dropwise to polyethylene glycol monomethyl ether-polyester diblock copolymer to obtain a mixed solution;

(B) adding ultrapure water dropwise to the mixed solution while stirring for the first time, and after continuing the second stirring, dialysis to remove the organic solvent and freeze-drying to obtain drug-loaded nanomicelles;

The polyethylene glycol monomethyl ether-polyester diblock copolymer is shown in formula (I),

Among them, -R- is or

m is the degree of polymerization, 10≤m≤900; n is the degree of polymerization, 10≤n≤420.

Preferably, in the step (A), the small molecule drug is methotrexate, 5-fluorouracil, cyclophosphamide, daunorubicin, doxorubicin, epirubicin, pirarubicin , camptothecins or taxanes.

Preferably, in the step (A), the concentration of the small molecule drug in the organic solvent is 0.1-10 mg/mL.

Preferably, in the step (A), the mass ratio of the small molecule drug to the polyethylene glycol monomethyl ether-polyester diblock copolymer is 0.01-1.

The invention provides a kind of preparation method of polyethylene glycol monomethyl ether-polyester diblock copolymer micelle, comprises the following steps:

Dissolve polyethylene glycol monomethyl ether-polyester diblock copolymer in an organic solvent, add ultrapure water dropwise while stirring for the first time, continue stirring for the second time, dialyze to remove the organic solvent and freeze-dry to obtain diblock copolymer micelles;

The polyethylene glycol monomethyl ether-polyester diblock copolymer is shown in formula (I),

Among them, -R- is or

m is the degree of polymerization, 10≤m≤900; n is the degree of polymerization, 10≤n≤420.

Preferably, the concentration of the polyethylene glycol monomethyl ether-polyester diblock copolymer in the organic solvent is 0.1-10 mg/mL.

Preferably, the speed of the first stirring is 100-2000 rpm.

Preferably, the rate of adding the ultrapure water dropwise is 0.05-5 mL/min.

Preferably, the volume ratio of the amount of ultrapure water to the amount of organic solvent is 0.01-20.

Preferably, the organic solvent is tetrahydrofuran, 1,4-dioxane, dimethylsulfoxide or N,N-dimethylformamide.

Compared with the prior art, the invention adopts the nano-sedimentation method to prepare the polyethylene glycol monomethyl ether-polyester diblock copolymer micelles or drug-loaded nano micelles. That is to dissolve polyethylene glycol monomethyl ether-polyester diblock copolymer in an organic solvent, add ultrapure water dropwise while stirring, and continue stirring to obtain nanomicelles; or mix a mixture of small molecule drugs and organic solvents Add it dropwise into the polyethylene glycol monomethyl ether block copolymer and stir to obtain a mixed solution. While stirring the mixed solution, add ultrapure water dropwise, and continue stirring to obtain drug-loaded nano micelles. The method prepares polyethylene glycol monomethyl ether-polyester diblock copolymer micelles, the operation is simple, the conditions are mild, and the generated nano-micelle particles can present a good monodisperse state, and the polyethylene glycol diblock copolymer prepared by the method Alcohol monomethyl ether-polyester diblock copolymer micelle core can encapsulate small molecule drugs with poor water solubility, and greatly improve the encapsulation efficiency and solubility of drugs. The obtained drug-loaded nano-micelles have a stable structure and small particle size. And easy to store.

Description of drawings

Fig. 1 is the nuclear magnetic resonance spectrum of the polyethylene glycol monomethyl ether-polyester diblock copolymer that embodiment 9 obtains in chloroform;

Fig. 2 is the nuclear magnetic resonance spectrum of the polyethylene glycol monomethyl ether-polyester diblock copolymer that embodiment 18 obtains in chloroform;

Fig. 3 is the transmission electron micrograph of the polyethylene glycol monomethyl ether-polyester diblock copolymer micelle prepared in embodiment 50;

Fig. 4 is the polyethylene glycol monomethyl ether-polyester diblock copolymer micelle size distribution figure that embodiment 50,62,86 obtains;

Fig. 5 is the graph of the impact of nanomicelles obtained in Examples 50, 62 and 86 of the present invention on the viability of MCF-7 cells;

Fig. 6 is the particle size distribution diagram of the drug-loaded nano-micelle prepared in Examples 91-93;

Fig. 7 is a graph showing the particle size distribution of drug-loaded nanomicelles prepared in Examples 91-93 within 3 weeks.

detailed description

In order to further understand the present invention, the preferred embodiments of the present invention are described below in conjunction with examples, but it should be understood that these descriptions are only to further illustrate the features and advantages of the present invention, rather than limiting the claims of the present invention.

The embodiment of the present invention discloses a preparation method of drug-loaded nano micelles, comprising the following steps:

(A) Add the mixture of small molecule drug and organic solvent dropwise to polyethylene glycol monomethyl ether-polyester diblock copolymer to obtain a mixed solution;

(B) adding ultrapure water dropwise to the mixed solution while stirring for the first time, and after continuing the second stirring, dialysis to remove the organic solvent and freeze-drying to obtain drug-loaded nanomicelles;

The polyethylene glycol monomethyl ether-polyester diblock copolymer is shown in formula (I),

Among them, -R- is or

m is the degree of polymerization, 10≤m≤900, preferably 20≤m≤800, more preferably 30≤m≤300; n is the degree of polymerization, 10≤n≤420, preferably 20≤n≤300, more preferably 40≤n≤250.

The present invention adopts the nano-sedimentation method to prepare the drug-loaded nano-micelle, which is simple to operate and has mild conditions. The inner core of the nano-micelle prepared by this method can wrap the small-molecule drug with poor water property, and greatly improve the encapsulation rate and solubility of the drug. The obtained drug-loaded nano micelles have a stable structure, a small particle size and are easy to store. The present invention preferably controls the particle size of the drug-loaded nanomicelles by changing the speed of adding ultrapure water, the amount of ultrapure water added, and the stirring speed in the preparation process, and adjusts the drug and polyethylene glycol monomethanone. The proportion of ether-polyester diblock copolymer is selected to select the best drug-loading conditions. In the invention, firstly, the mixture of the small molecule drug and the organic solvent is added dropwise into the polyethylene glycol monomethyl ether-polyester diblock copolymer to obtain a mixed solution. The finally formed micelles will encapsulate small molecule drugs inside, and the small molecule drugs are preferably hydrophobic antitumor drugs, more preferably methotrexate, 5-fluorouracil, cyclophosphamide, daunorubicin, Doxorubicin, epirubicin, pirarubicin, camptothecins or taxanes, most preferably 10-hydroxycamptothecin, doxorubicin or docetaxel. The organic solvent is an organic solvent miscible with water, preferably tetrahydrofuran, 1,4-dioxane, dimethyl sulfoxide or N,N-dimethylformamide, more preferably tetrahydrofuran. The polyethylene glycol monomethyl ether-polyester diblock copolymer is shown in formula (I),

Among them, -R- is or

m is the degree of polymerization, 10≤m≤900, preferably 20≤m≤800, more preferably 30≤m≤300; n is the degree of polymerization, 10≤n≤420, preferably 20≤n≤300, more preferably 40≤n≤250.

The polyethylene glycol monomethyl ether-polyester diblock copolymer is preferably obtained by polymerization of lactide or caprolactone and polyethylene glycol with a molecular weight of 500-10000; the lactide is D-lactide ester or L-lactide. Its preparation method is specifically:

Under anhydrous and oxygen-free conditions, add polyethylene glycol monomethyl ether and ester monomers into an ampoule, add a certain amount of toluene after azeotropic water removal, and the amount of toluene volume (mL) is the weight of ester monomers (g ), use a syringe to inject 0.1 mol/L stannous octoate toluene solution, the volume of stannous octoate and the molar ratio of ester monomers is 1/1000, and put it in an oil bath at 120°C for 24 hours. After the reaction is completed, settle with a large amount of ether, the ratio of ether to toluene is 10/1, filter with a Buchner funnel, dissolve the product in chloroform, settle with ether, filter with a Buchner funnel, and dry the product in a vacuum atmosphere for 24 hours. Acquired polyethylene glycol monomethyl ether-polyester diblock copolymer.

In the present invention, the mixture of the small molecule drug and the organic solvent is a homogeneous solution, preferably obtained by stirring for 4 to 6 hours, and the concentration of the small molecule drug in the organic solvent is preferably 0.1 to 10 mg/mL, more preferably 0.4 ~ 8mg/mL. The mixture of the small molecule drug and the organic solvent is added dropwise into the polyethylene glycol monomethyl ether-polyester diblock copolymer to obtain a mixed solution, and the small molecule drug and the polyethylene glycol monomethyl ether-polyester The mass ratio of the diblock copolymer is preferably 0.01 to 1, more preferably 0.05 to 0.5. Since the polyethylene glycol monomethyl ether-polyester diblock copolymer can be selected from ester monomers with different configurations in the preparation process, it can also be selected to have different configurations when preparing a mixed solution with the mixture. Polyethylene glycol monomethyl ether-polyester two-block copolymers with structural segments are mixed, and the left-handed and right-handed segments can interact with each other, so that they have stereocomplexity when forming micelles, thereby improving Drug encapsulation efficiency and solubility. The mass ratio of polyethylene glycol monomethyl ether-polyester diblock copolymers with different configuration segments is preferably 1:1. Preferably, after the mixture is added dropwise into the polyethylene glycol monomethyl ether-polyester diblock copolymer, stirring is performed; the stirring time is preferably 1-3 hours.

After the mixed solution is obtained, ultrapure water is added dropwise to the mixed solution while being stirred for the first time, and after the second stirring is continued, the organic solvent is removed by dialysis and freeze-dried to obtain drug-loaded nano micelles. The device for dropping the ultrapure water is preferably a syringe pump, and the speed of adding the ultrapure water is preferably 0.05-5 mL/min, more preferably 0.1-3 mL/min. The volume ratio of the amount of the ultrapure water to the amount of the organic solvent is preferably 0.01-20, more preferably 0.1-10. When the mixed solution is stirred for the first time, ultrapure water is added dropwise. The speed of the first stirring is preferably 100-2000 rpm, more preferably 800-1500 rpm. After the dropwise addition, the second stirring is continued to obtain the drug-loaded nano micelles, and the time of the second stirring is preferably 8-12 hours. The speed of the second stirring is preferably the same as that of the first stirring. In order to improve the purity of the obtained drug-loaded nanomicelles, it is preferable to perform dialysis in ultrapure water after the second stirring, the dialysis time is preferably 20 to 30 hours, and the water is changed more than 5 times, and the dialysis is preferably dialysis with a MWCO of 3500 bag. In order to facilitate preservation, the obtained drug-loaded nanomicelles can also be freeze-dried.

The invention discloses a preparation method of polyethylene glycol monomethyl ether-polyester diblock copolymer micelles, comprising the following steps:

Dissolve polyethylene glycol monomethyl ether-polyester diblock copolymer in an organic solvent, add ultrapure water dropwise while stirring for the first time, continue stirring for the second time, dialyze to remove the organic solvent and freeze-dry to obtain diblock copolymer micelles;

The polyethylene glycol monomethyl ether-polyester diblock copolymer is shown in formula (I),

Among them, -R- is or

m is the degree of polymerization, 10≤m≤900, preferably 20≤m≤800, more preferably 30≤m≤300; n is the degree of polymerization, 10≤n≤420, preferably 20≤n≤300, more preferably 40≤n≤250.

The invention adopts the nano-sedimentation method to prepare the polyethylene glycol monomethyl ether-polyester diblock copolymer micelle, which has simple operation and mild conditions.

In the present invention, polyethylene glycol monomethyl ether-polyester diblock copolymer is dissolved in an organic solvent, ultrapure water is added dropwise while stirring for the first time, and after the second stirring is continued, the organic solvent is removed by dialysis and frozen. Dry to obtain nanomicelles. The organic solvent is an organic solvent miscible with water, preferably tetrahydrofuran, 1,4-dioxane, dimethyl sulfoxide or N,N-dimethylformamide, more preferably tetrahydrofuran. The polyethylene glycol monomethyl ether-polyester diblock copolymer is preferably prepared according to the method described in the above technical solution. The concentration of the polyethylene glycol monomethyl ether-polyester diblock copolymer in the organic solvent is preferably 0.1-10 mg/mL, more preferably 2-8 mg/mL.

The device for dropping the ultrapure water is preferably a syringe pump, and the speed of adding the ultrapure water is preferably 0.05-5 mL/min, more preferably 0.1-3 mL/min. The volume ratio of the amount of the ultrapure water to the amount of the organic solvent is preferably 0.01-20, more preferably 0.1-10. The mixed liquid of polyethylene glycol monomethyl ether-polyester diblock copolymer is stirred for the first time while adding ultrapure water dropwise, the speed of the first stirring is preferably 100-2000rpm, more preferably 800～1500rpm. After the dropwise addition, the second stirring is continued to obtain polyethylene glycol monomethyl ether-polyester diblock copolymer micelles, and the time of the second stirring is preferably 8-12 hours. The speed of the second stirring is preferably the same as that of the first stirring. In order to improve the purity of the obtained polyethylene glycol monomethyl ether-polyester diblock copolymer micelles, it is preferable to carry out dialysis in ultrapure water after stirring for the second time, and the dialysis time is preferably 20 to 30 hours. For more than 5 times, the dialysis bag with MWCO of 3500 is preferred. In order to facilitate preservation, the obtained polyethylene glycol monomethyl ether-polyester diblock copolymer micelles can also be freeze-dried.

Utilize the particle diameter of the drug-loaded nano micelles that dynamic light scattering measures obtains, measure continuously for 3 weeks, the result shows that its particle diameter change trend is basically consistent, thus we can know that the drug-loaded nano micelles structure prepared by the method of the present invention Stablize.

The method of the present invention prepares polyethylene glycol monomethyl ether-polyester diblock copolymer micelles, the operation is simple, the conditions are mild, and the generated polyethylene glycol monomethyl ether-polyester diblock copolymer micelle particles It can present a good monodisperse state, and the polyethylene glycol monomethyl ether-polyester diblock copolymer micelle inner core prepared by the method of the present invention can wrap small molecule drugs with poor water solubility, and greatly improve the encapsulation efficiency of drugs and solubility, the obtained drug-loaded nano-micelle has a stable structure, a small particle size, and is easy to store.

In order to further understand the present invention, the preparation method of the polyethylene glycol monomethyl ether-polyester diblock copolymer micelle provided by the present invention and the preparation method of the drug-loaded nano-micelle are described below in conjunction with the examples, protection of the present invention The scope is not limited by the following examples.

Embodiment 1-5

Preparation of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers initiated by polyethylene glycol monomethyl ether with different number-average molecular weights

Weigh 1.67g, 3.33g, 6.67g, 16.67g, and 33.33g of polyethylene glycol monomethyl ether (MPEG) with molecular weights of 500, 1000, 2000, 5000, and 10,000, respectively, and put them into reaction flasks, and remove water by azeotropy. Add 12g of dextrolactide (DLA) in an anhydrous and oxygen-free environment, ventilate, the volume of toluene (mL) is 10 times the weight of ester monomer (g) 120mL, stannous octoate and ester monomer The molar ratio is 1/1000, inject 1 mL of 0.1 mol/L stannous octoate solution in toluene with a syringe, and put it in an oil bath at 120°C for 24 hours. After the reaction is complete, wait for the solution to cool down and settle with 1200mL of diethyl ether while stirring. The ratio of diethyl ether to toluene is 10/1. Filter the product through a Buchner funnel. Dissolve the product in chloroform and settle it with diethyl ether. Dry in a vacuum desiccator for 24 hours in a cold well to obtain polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers with different number average molecular weights.

Table 1 Number average molecular weight and reaction yield of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Examples 1-5

In Table 1, the number-average molecular weight M _is the number-average molecular weight of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers initiated by polyethylene glycol monomethyl ether of different number-average molecular weights, Measured by ¹ HNMR.

Embodiment 6-9

Preparation of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers with different degrees of polymerization initiated by polyethylene glycol monomethyl ether

Weigh 4g, 6g, 8g, and 12g of polyethylene glycol monomethyl ether (MPEG) with a molecular weight of 5000 and put them into a reaction bottle, remove water by azeotropic addition, and add dextrolactide (DLA) in an anhydrous and oxygen-free environment. ) 12g, ventilation, the volume of toluene (mL) is 10 times the weight of ester monomer (g) 120mL, the molar ratio of stannous octoate and ester monomer is 1/1000, inject 0.1mol/L octanoic acid with a syringe Put 1mL of stannous toluene solution in an oil bath at 120°C for 24h. After the reaction is complete, wait for the solution to cool down and settle with 1200mL of diethyl ether while stirring. The ratio of diethyl ether to toluene is 10/1. Filter the product through a Buchner funnel. Dissolve the product in chloroform and settle it with diethyl ether. Dry in a vacuum desiccator for 24 hours in a cold well to obtain polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers with different polymerization degrees initiated by polyethylene glycol monomethyl ether.

Table 2 Number average molecular weight and reaction yield of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Examples 6-9

In Table 2, the number-average molecular weight M _is the number-average molecular weight of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers with different degrees of polymerization initiated by polyethylene glycol monomethyl ether, which is represented by It was determined by ¹ H NMR.

See Figure 1 for the nuclear magnetic resonance spectrum of the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 9. Figure 1 shows the polyethylene glycol monomethyl ether obtained in Example 9. NMR spectrum of methyl ether-polyester diblock copolymer in chloroform. In Figure 1, 1.6ppm (S,3H,HO-CH(CH ₃ )OC(O)) 3.68ppm (S,2H,CH ₂ CH ₂ O)), 5.2ppm (S,3H,HO-CH(CH ₃ ) OC(O)). Figure 1 shows that the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 9 has the structure of formula (I).

Examples 10-14: Preparation of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers initiated by polyethylene glycol monomethyl ether with different number average molecular weights

Weigh 1.67g, 3.33g, 6.67g, 16.67g, and 33.33g of polyethylene glycol monomethyl ether (MPEG) with molecular weights of 500, 1000, 2000, 5000, and 10,000, respectively, and put them into reaction flasks, and remove water by azeotropy. Add 12g of L-lactide (LLA) in an anhydrous and oxygen-free environment, ventilate, the volume of toluene (mL) is 10 times the weight (g) of the ester monomer, that is, 120mL, the volume of stannous octoate and ester The molar ratio of the quasi-monomer is 1/1000, inject 1 mL of 0.1 mol/L stannous octoate toluene solution with a syringe, and put it in an oil bath at 120°C for 24 hours. After the reaction is complete, wait for the solution to cool down and settle with 1200mL of diethyl ether while stirring. The ratio of diethyl ether to toluene is 10/1. Filter the product through a Buchner funnel. Dissolve the product in chloroform and settle it with diethyl ether. Dry in a vacuum desiccator for 24 hours in a cold well to obtain polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers initiated by polyethylene glycol monomethyl ether with different number average molecular weights.

Table 3 Number average molecular weight and reaction yield of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Examples 10-14

In the above table, the number average molecular weight _Mn is the number average molecular weight of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer initiated by polyethylene glycol monomethyl ether with different number average molecular weights, Measured by ¹ HNMR.

Examples 15-18: Preparation of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers with different degrees of polymerization initiated by polyethylene glycol monomethyl ether

Weigh 4g, 6g, 8g, and 12g of polyethylene glycol monomethyl ether (MPEG) with a molecular weight of 5000, respectively, and put them into the reaction bottle, remove water by azeotropy, and add L-lactide (LLA) in an anhydrous and oxygen-free environment 12g, ventilation, the volume of toluene (mL) is 10 times the weight (g) of the ester monomer, that is, 120mL, the volume of stannous octoate and the molar ratio of the ester monomer is 1/1000, inject 0.1 with a syringe mol/L stannous octoate in toluene solution 1mL, put in 120℃ oil bath for 24h. After the reaction is complete, wait for the solution to cool down and settle with 1200mL of diethyl ether while stirring. The ratio of diethyl ether to toluene is 10/1. Filter the product through a Buchner funnel. Dissolve the product in chloroform and settle it with diethyl ether. Dry in a vacuum desiccator in a cold well for 24 hours to obtain polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers with different polymerization degrees initiated by polyethylene glycol monomethyl ether.

Table 4 Number average molecular weight and reaction yield of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Examples 15-17

In Table 4, the number-average molecular weight _Mn is the number-average molecular weight of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers with different degrees of polymerization initiated by polyethylene glycol monomethyl ether. It was determined by ¹ H NMR.

See Figure 2 for the nuclear magnetic resonance spectrum of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 18. Figure 2 shows the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer obtained in Example 18. NMR spectra of polyester diblock copolymers in chloroform. In Figure 2, 1.6ppm (S,3H,HO-CH(CH ₃ )OC(O)) 3.68ppm (S,2H,CH ₂ CH ₂ O)), 5.2ppm (S,3H,HO-CH(CH ₃ ) OC(O)). Figure 2 shows that the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 14 has the structure of formula (I).

Examples 19-23: Preparation of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymer initiated by polyethylene glycol monomethyl ether with different number average molecular weights

Weigh 1.32g, 2.63g, 5.26g, 13.16g, and 26.36g of polyethylene glycol monomethyl ether (MPEG) with molecular weights of 500, 1000, 2000, 5000, and 10,000, respectively, and put them into reaction flasks, and remove water with azeotropy. Add 12g of caprolactone in an anhydrous and oxygen-free environment, ventilate, the volume of toluene (mL) is 10 times the weight of caprolactone monomer (g) 120mL, the volume of stannous octoate and the mole of caprolactone monomer The ratio is 1/1000, inject 1 mL of 0.1 mol/L stannous octoate toluene solution with a syringe, and put it in an oil bath at 120°C for 24 hours. After the reaction is complete, wait for the solution to cool down and settle with 1200mL of diethyl ether while stirring. The ratio of diethyl ether to toluene is 10/1. Filter the product through a Buchner funnel. Dissolve the product in chloroform and settle it with diethyl ether. Dry in a vacuum desiccator for 24 hours in a cold well to obtain polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers with different number average molecular weights.

Table 5 Number average molecular weight and reaction yield of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers prepared in Examples 19-23

In Table 5, the number-average molecular weight M _n is the number-average molecular weight of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers with different number-average molecular weights, measured by ¹ H NMR.

Examples 24-27: Preparation of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers with different degrees of polymerization initiated by polyethylene glycol monomethyl ether

Weigh 4g, 6g, 8g, and 12g of polyethylene glycol monomethyl ether (MPEG) with a molecular weight of 5000, respectively, and put them into the reaction bottle, remove water by azeotropy, add 12g of caprolactone in an anhydrous and oxygen-free environment, and ventilate , the volume of toluene (mL) is 10 times the weight (g) of caprolactone monomer 120mL, the volume dosage of stannous octoate and the molar ratio of caprolactone monomer is 1/1000, inject 0.1mol/L octoate Tin toluene solution 1mL was placed in an oil bath at 120°C for 24h. After the reaction is complete, wait for the solution to cool down and settle with 1200mL of diethyl ether while stirring. The ratio of diethyl ether to toluene is 10/1. Filter the product through a Buchner funnel. Dissolve the product in chloroform and settle it with diethyl ether. Dry in a vacuum desiccator for 24 hours in a cold well to obtain a polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymer.

Table 6 Number average molecular weight and reaction yield of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers in Examples 24 to 27

In Table 6, the number-average molecular weight _Mn is the number-average molecular weight of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers with different polymerization degrees initiated by polyethylene glycol monomethyl ether. ¹ H NMR determined

Examples 28-30

Take 100 mg of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 18 and dissolve them in tetrahydrofuran, 1,4-dioxane, dimethyl sulfoxide and In N,N-dimethylformamide, the concentration is 2.5mg/mL, stir for 3h, set the flow rate of the syringe pump to 0.1mL/min, the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Stir the dissolved polyester block copolymer solution on a stirrer, and add Milli-Q dropwise to the solution at a constant speed with a set syringe pump. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q for 24 h with a dialysis bag with MWCO of 3500, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 31-33

Take 100 mg of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 18, and prepare tetrahydrofuran solutions with concentrations of 2 mg/mL, 2.5 mg/mL and 3 mg/mL respectively , stirred for 3 hours, set the syringe pump flow rate to 0.1mL/min, the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000r/min. Stir the dissolved polyester block copolymer solution on a stirrer, and add Milli-Q dropwise to the solution at a constant speed with a set syringe pump. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO: 3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 34-36

Take three parts of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 18, 100 mg for each part, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, Set the flow rate of the syringe pump to 0.1mL/min, the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 500rpm, 1000rpm, and 1500rpm respectively. Put the dissolved polyester lactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to drop Milli-Q into the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO: 3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 37-39

Take three parts of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 18, 100 mg for each part, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, Set the flow rate of the syringe pump to 0.1mL/min, the flow rate (mL) to 20mL, 25mL, and 30mL, respectively, and set the stirring speed of the stirrer to 1000rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO: 3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 40-42

Take three parts of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 18, 100 mg for each part, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, Set the flow rate of the syringe pump to 0.1mL/min, 0.3mL/min, and 0.5mL/min respectively, set the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO: 3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 43-47

Weigh 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Examples 1 to 5, dissolve them in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 hours, and set The flow rate of the syringe pump is 0.1 mL/min, the flow rate (mL) is set to 25 mL, and the stirring speed of the stirrer is set to 1000 rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 48-50

Weigh 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Examples 7 to 9, dissolve them in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 hours, and set The flow rates of the syringe pumps were 0.1 mL/min, 0.3 mL/min, and 0.5 mL/min, the flow rate (mL) was set to 25 mL, and the stirring speed of the stirrer was set to 1000 rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Fig. 3 is a transmission electron micrograph of the nanomicelle prepared in Example 50. It can be seen from Fig. 3 that the described method has prepared the nanomicelle.

Fig. 4 is the particle size distribution diagram of nano micelles obtained in Examples 50, 62, and 86, and the particle size distribution diagram is measured by dynamic light scattering (DLS). Figure 4, The dynamic hydrodynamic radius distribution of the nanomicelle obtained for embodiment 50, For the dynamic hydrodynamic radius distribution of the nanomicelle obtained in embodiment 62, Dynamic hydrodynamic radius distribution of nanomicelles obtained for Example 86.

Examples 51-53

Take three parts of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 5, each 100 mg, dissolve in tetrahydrofuran solution with a concentration of 2.5 mg/mL, stir for 3 h, Set the flow rate of the syringe pump to 0.1mL/min, 0.3mL/min, and 0.5mL/min respectively, set the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 54-58

Weigh 100 mg of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Examples 10-14, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 hours, and set The flow rate of the syringe pump is 0.1 mL/min, the flow rate (mL) is set to 25 mL, and the stirring speed of the stirrer is set to 1000 rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 59-62

Weigh 100 mg of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Examples 15-18, dissolve them in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 hours, and set The flow rate of the syringe pump is 0.1 mL/min, the flow rate (mL) is set to 25 mL, and the stirring speed of the stirrer is set to 1000 rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 63-65

Take three parts of polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 14, 100 mg for each part, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, Set the flow rate of the syringe pump to 0.1mL/min, 0.3mL/min, and 0.5mL/min, set the flow rate (mL) to 25mL, set the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 66-70

Weigh 100 mg of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers prepared in Examples 19-23, dissolve them in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 hours, and set The flow rate of the syringe pump is 0.1 mL/min, the flow rate (mL) is set to 25 mL, and the stirring speed of the stirrer is set to 1000 rpm. Put the dissolved polycaprolactone block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 71-74

Weigh 100 mg of the polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymers prepared in Examples 24 to 27, dissolve them in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 hours, and set The flow rate of the syringe pump is 0.1 mL/min, the flow rate (mL) is set to 25 mL, and the stirring speed of the stirrer is set to 1000 rpm. Put the dissolved polycaprolactone block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 75-77

Take three parts of polyethylene glycol monomethyl ether-poly(ε-caprolactone) diblock copolymer prepared in Example 23, 100 mg for each part, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, Set the flow rate of the syringe pump to 0.1mL/min, 0.3mL/min, and 0.5mL/min, set the flow rate (mL) to 25mL, set the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the dissolved polylactic acid block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 78-82

The polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Examples 1-5 and the polyethylene glycol monomethyl ether-poly(L-lactide) prepared in Examples 10-14 were mixed Mix 50 mg of lactide) two block copolymers, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, set the flow rate of the syringe pump to 0.1 mL/min, set the flow rate (mL) to 25 mL, and set the stirrer The stirring speed is 1000rpm. Put the dissolved polycaprolactone block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 83-86

The polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Examples 6-9 and the polyethylene glycol monomethyl ether-poly(L-lactide) prepared in Examples 15-18 Mix 50 mg of lactide) two block copolymers, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, set the flow rate of the syringe pump to 0.1 mL/min, set the flow rate (mL) to 25 mL, and set the stirrer The stirring speed is 1000rpm. Put the dissolved polycaprolactone block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Examples 87-89

The polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 5 was mixed with the polyethylene glycol monomethyl ether-poly(L-lactide) prepared in Example 14 Mix 50 mg of block copolymers, dissolve in tetrahydrofuran solution at a concentration of 2.5 mg/mL, stir for 3 h, set the flow rate of the syringe pump to 0.1 mL/min, 0.3 mL/min, 0.5 mL/min, and set the flow rate (mL) to Set as 25mL, set the stirring speed of the stirrer as 1000rpm. Put the dissolved polycaprolactone block copolymer solution on the stirrer to stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution was constant volume to obtain nano micelles.

Example 90

The toxicity test of the nano micelles that embodiment 50,62,86 obtains in MCF-7 cell:

The nanomicelles and PEI obtained in Examples 50, 62, and 86 were respectively configured into cell culture media with concentrations of 0.01 μg/mL, 0.025 μg/mL, 0.05 μg/mL, and 0.10 μg/mL,

First, MCF-7 cells were planted in a 96-well culture plate, and the culture medium was removed after 24 hours of culture. The experimental group was added the cell culture medium of nanomicelles obtained in the above-mentioned various concentrations of Examples 47, 57, and 79, and the control group was added with the above-mentioned Various concentrations of PEI. MTT was added after culturing for 72 hours, the culture medium was aspirated after 4 hours, and DMSO was added to measure the OD value at 490nm wavelength of the control group and the experimental group on a microplate reader.

The experimental results are shown in Fig. 5. Fig. 5 is a graph showing the effect of nanomicelles obtained in Examples 50, 62, and 86 of the present invention on the survival rate of MCF-7 cells. In Fig. 5, The influence curve of the nanomicelle obtained for the embodiment 50 of different concentrations on the cell viability, The influence curve of the nanomicelle obtained for the embodiment 62 of different concentrations on the cell viability, The influence curve of the nano micelles that the embodiment 86 of different concentrations obtains on cell viability; Effect curve of different concentrations of PEI on cell viability.

As shown in Figure 5, in all tested concentration ranges, the cell survival rate of the nanomicelles with the highest concentration of 0.1 mg L ^-1 was above 80% after 72 h of culture. It can be seen that the nanomicelle has low cytotoxicity and can be safely used as a biocompatible carrier for transporting biologically active substances.

Examples 91-93

Ethylene glycol monomethyl ether-polyester diblock copolymer 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 4 was selected respectively, and the poly(D-lactide) diblock copolymer prepared in Example 13 Ethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 4 and implementation Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 13.

Dissolve 10-hydroxycamptothecin in tetrahydrofuran, the concentration of 10-hydroxycamptothecin is 0.4mg/mL, stir for 5h, after dissolving, add 10-hydroxycamptothecin-tetrahydrofuran solution dropwise to the weighed poly The solution that the mass ratio of 10-hydroxycamptothecin and polyethylene glycol monomethyl ether-polyester diblock copolymer is configured into 1:5 is continued to stir 2h, set the flow rate of the syringe pump to 0.1mL/min, the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved with 10-hydroxycamptothecin on the stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

After calculation, the encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 91 is 92%, and the drug-loading efficiency is 12.2%; the encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 92 is 100%, and the drug-loading efficiency is 13.2%; the encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 93 was 100%, and the drug-loading efficiency was 13.2%.

Fig. 6 is a particle size distribution diagram of drug-loaded nanomicelles prepared in Examples 91-93, and the particle size distribution diagram is measured by dynamic light scattering (DLS). Figure 6, For the dynamic hydrodynamic radius distribution of the drug-loaded nanomicelle obtained in Example 91, For the dynamic hydrodynamic radius distribution of the drug-loaded nanomicelle obtained in Example 92, The dynamic hydrodynamic radius distribution of the drug-loaded nanomicelle obtained in Example 93.

Fig. 7 is the particle size distribution diagram of the drug-loaded nanomicelles prepared in Examples 91-93 within 3 weeks, a is the particle size distribution of the drug-loaded nanomicelles prepared in Examples 91-93 in 1 week, b c is the particle size distribution of the drug-loaded nanomicelles prepared in Examples 91-93 in 1 week, c is the particle size distribution of the drug-loaded nanomicelles prepared in Examples 91-93, For the dynamic hydrodynamic radius distribution of the drug-loaded nanomicelle obtained in Example 91, For the dynamic hydrodynamic radius distribution of the drug-loaded nanomicelle obtained in Example 92, The dynamic hydrodynamic radius distribution of the drug-loaded nanomicelle obtained in Example 93.

It can be seen from Figure 7 that the drug-loaded nanomicelles prepared by the present invention have good stability.

Examples 94-96

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 5, prepared in Example 14 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 5 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 14.

Dissolve 10-hydroxycamptothecin in tetrahydrofuran, the concentration of 10-hydroxycamptothecin is 0.4mg/mL, stir for 5h, after dissolving, add 10-hydroxycamptothecin-tetrahydrofuran solution dropwise to the weighed poly Prepare a solution of 10-hydroxycamptothecin and polyethylene glycol monomethyl ether-polyester diblock copolymer with a mass ratio of 1:5 in the ester block copolymer and continue to stir for 2 hours, set the flow rate of the syringe pump to 0.1mL/min , The flow rate (mL) is set to 25 mL, and the stirring speed of the stirrer is set to 1000 rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved with 10-hydroxycamptothecin on the stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 94 was 10.16%, and the drug-loading efficiency was 75.41%; the encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 96 was 11.92%, and the drug-loading efficiency was 90.29%; The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 96 was 12.95%, and the drug-loading efficiency was 99.22%.

Examples 97-101

Weigh 100 mg of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer prepared in Example 15, respectively. Dissolve 10-hydroxycamptothecin in tetrahydrofuran, the concentration of 10-hydroxycamptothecin is 0.4mg/mL, stir for 5h, after dissolving, add 10-hydroxycamptothecin-tetrahydrofuran solution dropwise to the weighed poly The mass ratio of 10-hydroxycamptothecin to polyethylene glycol monomethyl ether-polyester diblock copolymer in the ester block copolymer is 1:20, 1:10, 3:20, 4:20, Continue to stir the 5:20 solution for 2 hours, set the flow rate of the syringe pump to 0.1mL/min, set the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved with 10-hydroxycamptothecin on the stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 97 was 10.95%, and the drug-loading efficiency was 73.61%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 98 was 11.7%, and the drug-loading efficiency was 80.3%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 99 was 13.1%, and the drug-loading efficiency was 100%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 100 was 12.1%, and the drug-loading efficiency was 90.21%;

The encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 101 was 11.2%, and the drug-loading efficiency was 78.31%.

Examples 102-106

Weigh 100 mg of the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6, respectively. Dissolve 10-hydroxycamptothecin in tetrahydrofuran, the concentration of 10-hydroxycamptothecin is 0.4mg/mL, stir for 5h, after dissolving, add 10-hydroxycamptothecin-tetrahydrofuran solution dropwise to the weighed poly The mass ratio of 10-hydroxycamptothecin to polyethylene glycol monomethyl ether-polyester diblock copolymer is 1:20 and 1:10 respectively in the ethylene glycol monomethyl ether-polyester diblock copolymer , The solution of 3:20, 4:20, and 5:20 was continuously stirred for 2 hours, the flow rate of the syringe pump was set to 0.1mL/min, the flow rate (mL) was set to 25mL, and the stirring speed of the stirrer was set to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved with 10-hydroxycamptothecin on the stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 102 was 11.03%, and the drug-loading efficiency was 75.22%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 103 was 11.98%, and the drug-loading efficiency was 81.21%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 104 was 12.94%, and the drug-loading efficiency was 98.4%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 105 was 12.29%, and the drug-loading efficiency was 90.18%;

The encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 106 was 11.03%, and the drug-loading efficiency was 76.94%.

Examples 107-111

The polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 was mixed with the polyethylene glycol monomethyl ether-poly(L-lactide) prepared in Example 15 50 mg of each block copolymer was mixed to prepare 5 parts of the same mixed polyurethane block copolymer. Dissolve 10-hydroxycamptothecin in tetrahydrofuran, the concentration of 10-hydroxycamptothecin is 0.4mg/mL, stir for 5h, after dissolving, add 10-hydroxycamptothecin-tetrahydrofuran solution dropwise to the weighed poly The mass ratio of 10-hydroxycamptothecin to polyethylene glycol monomethyl ether-polyester diblock copolymer is 1:20 and 1:10 respectively in the ethylene glycol monomethyl ether-polyester diblock copolymer , The solution of 3:20, 4:20, and 5:20 was continuously stirred for 2 hours, the flow rate of the syringe pump was set to 0.1mL/min, the flow rate (mL) was set to 25mL, and the stirring speed of the stirrer was set to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved with 10-hydroxycamptothecin on the stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 107 was 11.93%, and the drug-loading efficiency was 78.26%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 108 was 12.24%, and the drug-loading efficiency was 85.21%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 109 was 12.93%, and the drug-loading efficiency was 98.64%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 110 was 12.31%, and the drug-loading efficiency was 83.51%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 111 was 11.81%, and the drug-loading efficiency was 79.15%.

Examples 112-114

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve methotrexate in tetrahydrofuran, the concentration of methotrexate is 0.4mg/mL, stir for 5h, after dissolving, add methotrexate-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether - The polyester diblock copolymer is configured so that the mass ratio of methotrexate to polyethylene glycol monomethyl ether-polyester diblock copolymer is 1:20, 1:10, 3:20, 4:20, respectively , The solution of 5:20 was stirred continuously for 2h, the flow rate of the syringe pump was set to 0.1mL/min, the flow rate (mL) was set to 25mL, and the stirring speed of the stirrer was set to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved in methotrexate on a stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 112 was 21.42%, and the drug-loading efficiency was 82.2%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 113 was 21.38%, and the drug-loading efficiency was 83.36%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 114 was 22.93%, and the drug-loading efficiency was 95.83%.

Examples 115-117

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve cyclophosphamide in tetrahydrofuran, the concentration of cyclophosphamide is 0.4mg/mL, stir for 5h, after dissolving, add cyclophosphamide-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether-polyester The mass ratio of cyclophosphamide and polyethylene glycol monomethyl ether-polyester diblock copolymer in the diblock copolymer is 1:20, 1:10, 3:20, 4:20, 5:20, respectively Continue to stir the solution for 2 h, set the flow rate of the syringe pump to 0.1 mL/min, set the flow rate (mL) to 25 mL, and set the stirring speed of the stirrer to 1000 rpm. Put the cyclophosphamide-dissolved polyethylene glycol monomethyl ether-polyester diblock copolymer solution on a stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 115 was 51.57%, and the drug-loading efficiency was 73.41%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 116 was 51.94%, and the drug-loading efficiency was 75.94%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 117 was 60.21%, and the drug-loading efficiency was 89.34%.

Examples 118-120

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve 5-fluorouracil in tetrahydrofuran, the concentration of 5-fluorouracil is 0.4mg/mL, stir for 5h, after dissolving, add 5-fluorouracil-tetrahydrofuran solution dropwise to the weighed polyethylene glycol The monomethyl ether-polyester diblock copolymer is configured so that the mass ratios of 5-fluorouracil and polyethylene glycol monomethyl ether-polyester diblock copolymer are 1:20, 1:10, and 3:20, respectively , The 4:20, 5:20 solution continued to stir for 2h, set the flow rate of the syringe pump to 0.1mL/min, the flow rate (mL) to 25mL, and set the stirring speed of the stirrer to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved with 5-fluorouracil on a stirrer and stir, and add Milli-Q dropwise to the solution at a constant speed with a set syringe pump. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 118 was 42.12%, and the drug-loading efficiency was 83.56%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 119 was 43.72%, and the drug-loading efficiency was 82.13%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 120 was 50.92%, and the drug-loading efficiency was 98.27%.

Examples 121-123

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve docetaxel in tetrahydrofuran, the concentration of docetaxel is 0.4mg/mL, stir for 5h, after dissolving, add the docetaxel-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether-polyester The mass ratios of docetaxel and polyethylene glycol monomethyl ether-polyester diblock copolymer in the diblock copolymer are 1:20, 1:10, 3:20, 4:20, 5:20, respectively Continue to stir the solution for 2 h, set the flow rate of the syringe pump to 0.1 mL/min, set the flow rate (mL) to 25 mL, and set the stirring speed of the stirrer to 1000 rpm. The docetaxel-dissolved polyethylene glycol monomethyl ether-polyester diblock copolymer solution was placed on a stirrer and stirred, and Milli-Q was added dropwise to the solution at a constant speed with a set syringe pump. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 121 was 14.85%, and the drug-loading efficiency was 83.56%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 122 was 14.37%, and the drug-loading efficiency was 84.67%;

The encapsulation efficiency of the drug-loaded nanomicelles obtained in Example 123 was 15.93%, and the drug-loading efficiency was 95.33%.

Examples 124-126

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve daunorubicin in tetrahydrofuran, the concentration of daunorubicin is 0.4mg/mL, stir for 5h, after dissolving, add daunorubicin-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether -The mass ratio of daunorubicin and polyethylene glycol monomethyl ether-polyester diblock copolymer in the polyester diblock copolymer is 1:20, 1:10, 3:20, 4:20 respectively , The solution of 5:20 was stirred continuously for 2h, the flow rate of the syringe pump was set to 0.1mL/min, the flow rate (mL) was set to 25mL, and the stirring speed of the stirrer was set to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved in daunorubicin on the stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 124 was 14.33%, and the drug-loading efficiency was 88.49%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 125 was 14.41%, and the drug-loading efficiency was 86.27%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 126 was 15.86%, and the drug-loading efficiency was 96.27%.

Examples 127-129

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve doxorubicin in tetrahydrofuran, the concentration of doxorubicin is 0.4mg/mL, stir for 5h, after dissolving, add doxorubicin-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether-polyester The mass ratios of doxorubicin and polyethylene glycol monomethyl ether-polyester diblock copolymer in the diblock copolymer are 1:20, 1:10, 3:20, 4:20, 5:20, respectively Continue to stir the solution for 2 h, set the flow rate of the syringe pump to 0.1 mL/min, set the flow rate (mL) to 25 mL, and set the stirring speed of the stirrer to 1000 rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved in doxorubicin on a stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 127 was 14.68%, and the drug-loading efficiency was 86.12%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 128 was 14.52%, and the drug-loading efficiency was 85.93%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 129 was 15.84%, and the drug-loading efficiency was 96.31%.

Examples 130-132

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve epirubicin in tetrahydrofuran, the concentration of epirubicin is 0.4mg/mL, stir for 5h, after dissolving, add epirubicin-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether -The mass ratio of epirubicin and polyethylene glycol monomethyl ether-polyester diblock copolymer in the polyester diblock copolymer is 1:20, 1:10, 3:20, 4:20 respectively , The solution of 5:20 was stirred continuously for 2h, the flow rate of the syringe pump was set to 0.1mL/min, the flow rate (mL) was set to 25mL, and the stirring speed of the stirrer was set to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved in epirubicin on a stirrer and stir, and drop Milli-Q into the solution at a constant speed with a set syringe pump. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 130 was 14.77%, and the drug-loading efficiency was 85.35%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 131 was 14.64%, and the drug-loading efficiency was 84.95%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 132 was 16.03%, and the drug-loading efficiency was 96.12%.

Examples 133-135

Polyethylene glycol monomethyl ether-polyester diblock copolymers were respectively selected from 100 mg of polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymers prepared in Example 6, prepared in Example 15 Polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymer 100 mg, the polyethylene glycol monomethyl ether-poly(D-lactide) diblock copolymer prepared in Example 6 and Mix 50 mg each of the polyethylene glycol monomethyl ether-poly(L-lactide) diblock copolymers prepared in Example 15.

Dissolve pirarubicin in tetrahydrofuran, the concentration of pirarubicin is 0.4mg/mL, stir for 5h, after dissolving, add the pirarubicin-tetrahydrofuran solution dropwise to the weighed polyethylene glycol monomethyl ether - The polyester diblock copolymer is configured so that the mass ratio of pirarubicin to polyethylene glycol monomethyl ether-polyester diblock copolymer is 1:20, 1:10, 3:20, 4:20, respectively , The solution of 5:20 was stirred continuously for 2h, the flow rate of the syringe pump was set to 0.1mL/min, the flow rate (mL) was set to 25mL, and the stirring speed of the stirrer was set to 1000rpm. Put the polyethylene glycol monomethyl ether-polyester diblock copolymer solution dissolved in pirarubicin on a stirrer and stir, and use the set syringe pump to add Milli-Q dropwise to the solution at a constant speed. Continue to stir for 10 h after the dropwise addition, dialyze in Milli-Q with a dialysis bag (MWCO=3500) for 24 h, and change the water more than 5 times. After dialysis, the solution is freeze-dried to obtain biologically active drug-loaded nano-micelles.

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 133 was 14.91%, and the drug-loading efficiency was 84.23%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 134 was 14.93%, and the drug-loading efficiency was 84.16%;

The encapsulation efficiency of the drug-loaded nanomicelle obtained in Example 135 was 16.12%, and the drug-loading efficiency was 96.32%.

The above description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the invention. The general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Accordingly, the present invention shall not be limited to the Examples shown herein.

Claims (10)

1. a kind of preparation method of medicament-carried nano micelle, comprises the following steps：

(A) mixture of small-molecule drug and organic solvent is added drop-wise in poly glycol monomethyl ether-polyester biblock copolymer, Obtain mixed solution；

(B) Deca ultra-pure water while carrying out stirring for the first time by the mixed solution, after continuing second stirring, dialysis removing Simultaneously lyophilizing obtains medicament-carried nano micelle to organic solvent；

Shown in the poly glycol monomethyl ether-polyester biblock copolymer such as formula (I),

Wherein ,-R- is

M is the degree of polymerization, 10≤m≤900；N is the degree of polymerization, 10≤n≤420；

Poly glycol monomethyl ether-the polyester biblock copolymer includes the poly glycol monomethyl ether of various configuration segment-poly- Ester di-block copolymer；

Mass ratio between the poly glycol monomethyl ether-polyester biblock copolymer of the various configuration segment is 1:1.

2. preparation method according to claim 1, it is characterised in that in the step (A), the small-molecule drug is first Aminopterin, 5-fluorouracil, cyclophosphamide, daunorubicin, amycin, epirubicin, Pirarubicin, camptothecin or Ramulus et folium taxi cuspidatae Class.

3. preparation method according to claim 1, it is characterised in that in the step (A), the small-molecule drug is having Concentration in machine solvent is 0.1～10mg/mL.

4. preparation method according to claim 1, it is characterised in that in the step (A), the small-molecule drug with it is poly- The mass ratio of glycol monoethyl ether-polyester biblock copolymer is 0.01～1.

5. a kind of preparation method of poly glycol monomethyl ether-polyester biblock copolymer micelle, comprises the following steps：

Poly glycol monomethyl ether-polyester biblock copolymer is dissolved in organic solvent, carries out being dripped while stirring for the first time Plus ultra-pure water, after continuing second stirring, dialysis removes organic solvent and lyophilizing obtains di-block copolymer micelle；

Shown in the poly glycol monomethyl ether-polyester biblock copolymer such as formula (I),

Wherein ,-R- is

M is the degree of polymerization, 10≤m≤900；N is the degree of polymerization, 10≤n≤420；

Poly glycol monomethyl ether-the polyester biblock copolymer includes the poly glycol monomethyl ether of various configuration segment-poly- Ester di-block copolymer；

Mass ratio between the poly glycol monomethyl ether-polyester biblock copolymer of the various configuration segment is 1:1.

6. preparation method according to claim 5, it is characterised in that the block of poly glycol monomethyl ether-polyester two is total to Polymers concentration in organic solvent is 0.1～10mg/mL.

7. the preparation method according to any one of claim 1 or 5, it is characterised in that the speed of the first time stirring For 100～2000rpm.

8. the preparation method according to any one of claim 1 or 5, it is characterised in that the speed of the Deca ultra-pure water For 0.05～5mL/min.

9. the preparation method according to any one of claim 1 or 5, it is characterised in that the consumption of the ultra-pure water with have The volume ratio of machine solvent load is 0.01～20.

10. the preparation method according to any one of claim 1 or 5, it is characterised in that the organic solvent is tetrahydrochysene furan Mutter, 1,4- dioxane, dimethyl sulfoxide or N,N-dimethylformamide.